CN109287623A - A kind of purple U.S. pomegranate pollen storage practice - Google Patents
A kind of purple U.S. pomegranate pollen storage practice Download PDFInfo
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- CN109287623A CN109287623A CN201811214967.4A CN201811214967A CN109287623A CN 109287623 A CN109287623 A CN 109287623A CN 201811214967 A CN201811214967 A CN 201811214967A CN 109287623 A CN109287623 A CN 109287623A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N3/00—Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves or protection against physical influences such as UV radiation using chemical compositions; Grafting wax
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Abstract
The present invention provides a kind of purple U.S. pomegranate pollen storage practice, belongs to plant developmental biology field.Purple U.S. pomegranate pollen storage practice of the invention, comprising the following steps: pollen collecting, pollen is dry, pollen storage.The present invention forms pomegranate pollen collecting, drying, preservation and viability examination system for the first time, operating method is simple, repeatability is strong, effectively extend pomegranate pollen shelf-life, and keep the preferable vigor of storage period pollen, improves the success rate of pomegranate artificial hybridization pollination.
Description
Technical field
The present invention relates to plant developmental biology fields, and in particular to a kind of purple U.S. pomegranate pollen storage practice.
Background technique
Crossbreeding is the important channel of pomegranate new varieties initiative.Pollen viability is to ensure that crossbreeding successful one
Essential condition.Due to China the North and South cultivation pomegranate florescence it is inconsistent, Sichuan, Yunnan Province full-bloom stage in four months
Ten days, and the pomegranate full-bloom stage of Henan and Shandong District in may in, the last ten-days period, this brings one for the North and South different cultivars intermolecular hybrid
Fixed puzzlement.Pollen collection and storage are urgent problems to be solved during pomegranate Germplasm enhancement.Pomegranate pollen is store at present
Silica dehydrator, vetrifying solution PVS are mostly used when hiding4Etc. complex drying means, complex disposal process or undried
The method directly stored is handled, the storage time that will lead within the scope of the effective viability of pollen greatly shortens.Room temperature item
Under part, viability is lost completely within pomegranate pollen storage 5 days；Using liquid nitrogen storage pollen, later period pollen thaws and low temperature is multiple
Pollen activity can sharply decline after Soviet Union, with the first-class equal uncomfortable best storage practice for for pomegranate cross experiment pollen.This hair
It is bright using ' purple beauty ' pomegranate kind as test material, it is intended to study pomegranate pollen storage optimum treatment mode and holding conditions, it is miscellaneous for pomegranate
Breeding work is handed over to provide technical support.
' purple beauty ' pomegranate introduces Panzhihua City from Israel, through Panzhihua Agriculture and Forestry Science Research Institute, Goats in Liangshan Prefecture within 1996
Subtropical crop research institute, Horticultural Research Institute, Sichuan Academy of Agricultural Sciences combine the excellent variety of many years experiment sieving.' purple beauty '
Pomegranate tree-shaped half-open, tree vigo(u)r is strong, strong at principal stresses；Blade oblong, obovate, spire light red, old leaf peony；Sepal
Peony, holder peony, tubular or mitriform；Total flower amount is medium, and hermaphrodite flower ratio is higher.In Panxi Area, Sichuan Province subtropical zone gas
Under the conditions of time, ' purple beauty ' pomegranate is in rudiment mid-February, and March, early and middle ten days were buddingged, and the florescence is from late March to the first tenday period of a month in May；Under in September
Ten days fruit maturation, fruit be subsphaeroidal, pericarp peony, 587 grams of mean fruit weight.' purple beauty ' pomegranate keeping quality is strong, pericarp matter
Ground is coarse, and 3 millimeters of peel thickness；Seed aubergine, juice atropurpureus, seed spy are soft.' purple beauty ' pomegranate has pericarp dark red, outside
Beauty is seen, seed is softer, and commodity value is higher, great market potential.
Pomegranate pollen has to pass through viability test, can just determine whether to be used for hybridization pollination.Have to no determination
Pollen without viability and viability size, if blindly use, may by will lead to crossing work failure or can not be to miscellaneous
Knot fruit is reliably analyzed.The present invention includes the pomegranate pollen storage practice of three key technology steps, the technical operation
It is simple and easy, storage period pomegranate pollen activity is effectively maintained, is had in fields such as pomegranate production, breeding, scientific researches important
Application value.
Summary of the invention
For the deficiency of the prior art and research, the object of the present invention is to provide a kind of purple U.S. pomegranate pollen storage practice,
This method is simple,.
In order to achieve the above object, the present invention adopts the following technical scheme:
A kind of purple U.S. pomegranate pollen storage practice, comprising the following steps:
(1) it pollen collecting: when pomegranate flower is in full-bloom stage, reaches maturity in morning 8:00-10:00 acquisition, desire in bud
The flower put guarantees the validity of development of floral organs；
(2) pollen is dry: by the careful removal of petal of the flower of acquisition, holding chamber is interior to be spontaneously dried；To anther dehiscence loose powder
When, flower is gently vibrated, pollen is equably collected on clean blank sheet of paper, continues to dry in the air dry to pollen, drying regime is colored
Powder scatters completely, not at bulk；
(3) pollen storage: glass desicator internal washing is clean, and with 75% alcohol wipe inner wall, by the silica gel that changes colour
It pours into and is covered with drier bottom；Dried pomegranate pollen is fitted into container, seals, is placed in the drier of sealing,
It is saved in 4 DEG C.
Further technical solution, in the step (1), full-bloom stage is the four months bottoms in Sichuan province to early May, 70%~
75% purple U.S. pomegranate flower open period.
Further technical solution, in the step (1), the flower reached maturity be flower ovary obviously expand, flower pattern
For calabash shaped or the flower of tubular, selected flower is complete flower.
Further technical solution, in the step (2), natural drying be under 25 DEG C of Indoor Natural illumination it is dry for 24 hours.
Further technical solution, in the step (3), container is 10mL centrifuge tube, and preservation is placed in fridge freshness retaining layer 4
DEG C storage.
The present invention is using the purple U.S. pomegranate pollen vigor condition of fluid nutrient medium culture assay storage period.Pollen with dissipate
It broadcasts on liquid medium, makes its pollen germination, to measure a kind of method of pollen viability.Concrete operations are as follows: drop 1-2 drop
Liquid Culture is based on glass slide, takes a small amount of pollen to be uniformly sprinkling upon on fluid nutrient medium with pin, fluid nutrient medium is sucrose.
Glass slide is placed in the culture dish of humidity 85%-90%, dark culture 12h under the conditions of 25 DEG C.The normal pollen germination of viability
When rounded, pollen tube linear extension；Its pollen tube is bent after the pollen germination of viability difference；Do not have viable pollen grain not
It can sprout.
The invention has the following advantages:
1. pomegranate pollen collection method of the present invention, chooses the complete flower to reach maturity, guarantee the validity of development of floral organs,
The consistency for guaranteeing pollen fertility under natural conditions, avoids leading to pollen activity because different type development of floral organs degree is inconsistent
Difference problem.
2. pomegranate flower dried bean noodles drying method of the present invention, spontaneously dries indoors, compared in previous direct sunlight or drier,
Guarantee the objectivity of pollen activity and research under natural conditions.
3. pomegranate pollen storage practice of the present invention is placed multiple temperature gradients respectively and is stored, examined by pollen activity
It surveys, after a week, pollen at room temperature has completely lost viability.And 4 DEG C of preservations are used, storage pollen can be used directly
In experimental study, while avoiding in the prior art using the resuscitation process after -20 DEG C, -80 DEG C, the low-temperature treatments such as liquid nitrogen to pollen
Damage the problems such as, and this method is simple, repeatability is strong.
4. when pomegranate pollen vitality test of the present invention, the relative humidity of pollen cultures environment is 85%-90%, guarantee pollen
Sprout required moisture in incubation.
5. the present invention uses liquid MS for minimal medium, under the premise of fundamental component is constant, sucrose and grape are set
Two processing of sugar are measured by pollen activity and are found under identical storage time and holding conditions, and dextrose culture-medium pollen is living
Power is significantly lower than sucrose, illustrates that sucrose is that the fluid nutrient medium of sugar source is more suitable for purple U.S. pomegranate pollen in vitro culture.
6. the present invention forms the system of pomegranate pollen collecting, drying, preservation and vitality test for the first time, method has operation letter
The features such as single, rapid and repeatable strong, while this method extends the effective storage time of pomegranate pollen, and pomegranate people can be improved
The success rate of work hybridization pollination guarantees that crossbreeding is the main path of pomegranate breeding improved Varieties.
Detailed description of the invention
The drying and collection of Fig. 1 pollen
Fig. 2 pollen storage
Fig. 3 pollen activity determination step
Influence of Fig. 4 difference culture solution to pomegranate pollen germination rate
Fig. 5 different storage time, the variation diagram of purple U.S. pomegranate pollen germination rate.
Specific embodiment
To keep the purpose, technical solution and main points of the embodiment of the present invention clearer, below with reference to the embodiment of the present invention and
Attached drawing technical solution in the embodiment of the present invention is described in detail.
Embodiment
A kind of purple U.S. pomegranate pollen storage practice, comprising the following steps:
(1) it pollen collecting: when pomegranate flower is in full-bloom stage, reaches maturity in morning 8:00-10:00 acquisition, desire in bud
The flower put guarantees the validity of development of floral organs；Full-bloom stage is that the four months bottoms in Sichuan province are purple to early May, 70%~75%
U.S. pomegranate flower open period.The flower reached maturity is the flower that flower ovary is obviously expanded, flower pattern is calabash shaped or tubular, institute
Choosing flower is complete flower.
(2) pollen is dry: by the careful removal of petal of the flower of acquisition, holding chamber is interior to be spontaneously dried, and natural drying is in 25
Under DEG C Indoor Natural illumination it is dry for 24 hours；When anther dehiscence loose powder, flower is gently vibrated, pollen is equably collected clean
Blank sheet of paper on, continue to dry in the air dry to pollen, drying regime is that pollen scatters completely, not at bulk；
(3) pollen storage: glass desicator internal washing is clean, and with 75% alcohol wipe inner wall, by the silica gel that changes colour
(a kind of granulated dried agent, blue, bleach after water suction) pours into and is covered with drier bottom；Dried pomegranate pollen is packed into
In container, sealing is placed in the drier of sealing, is saved in 4 DEG C.Container is 10mL centrifuge tube, and preservation is placed in refrigerator guarantor
4 DEG C of fresh layer storages.
The dry multiple temperature gradients of pollen dispensed are stored.Pomegranate pollen after natural drying is in lower centrifuge tube
It inside saves at room temperature, pollen loses vigor completely after a week.When pollen storage temperature is 25 DEG C or is higher than the temperature, pollen is lost
Water is serious, influences later period germination rate；When reserve temperature is in 0 DEG C hereinafter, the solidification of intracellular Free water, pollen cell structure by
It destroys, pollen is made to lose viability.Though the use of the antifreezing agents such as glycerol protects cells from low temperature injury to a certain extent,
Low temperature can still cause pollen inevitably to injure, after cryopreservation the recovery of pollen and etc. make test become many and diverse,
Compared by multiple temperature, pollen activity is up to 42.1% under the conditions of 4 DEG C after a week for discovery storage, and experimental result is optimal, can
Effectively avoid problem above and drawback.
It with sugar source is respectively two kinds of liquid of sucrose and glucose when culture by its viability of the pomegranate flower powder periodic detection of storage
Body culture medium.1-2 drop of liquid culture medium is dripped on glass slide, takes a small amount of pollen to be placed in solution with pin.Pollen is uniform
It is sprinkling upon on fluid nutrient medium.Glass slide is placed in the culture dish of humidity 85%-90%, is placed in incubator and is trained for 25 DEG C
It supports (see Fig. 3).The quantity of statistics germination pollen grain, each repetition microscopy 3 or more are representative under an optical microscope after 12h
The visual field, calculate pollen germination rate: pollen germination rate (%)=pollen tube length be greater than pollen diameter pollen quantity/visual field in
Pollen frequence × 100%.
Under identical processing and holding conditions, two kinds of culture solutions pair of sucrose and glucose are added respectively in MS by comparing
The influence of pollen germination finds that the pollen germination rate of glucose (40.7%) culture is significantly lower than sucrose (46.2%) (Fig. 4).
It is tested and is found by multiple groups, MS+4.2g/L sucrose, germination rate is best, energy source equally is provided for pollen cell as carbon source,
Sucrose in water ratio glucose can preferably adjust the osmotic pressure in culture medium, and glucose easily makes cell dehydration as carbon source and grows not
It is good.As known from the above, pomegranate pollen vitality test is suitble to sucrose culture solution culture pomegranate pollen germination.
By data analysis found that method of the invention is particularly suitable for the short term storage of purple U.S. pomegranate pollen, in storage 21
After it, germination rate remains at 32.48%, within the scope of the effective germination rate of pollen (> 20.0%), when storing 130 days at 4 DEG C
Germination rate is still up to 21.5%.Illustrate that the storage practice can effectively maintain pomegranate pollen activity.The present invention has stored three
The pomegranate pollen of the moon still can be used for crossing work (Fig. 5), meet the crossbreeding of China pomegranate anniversary different regions pomegranate
And related scientific research work.
Claims (7)
1. a kind of purple U.S. pomegranate pollen storage practice, which comprises the following steps:
(1) pollen collecting: when pomegranate flower is in full-bloom stage, in morning 8:00-10:00 acquisition is reached maturity, a bud just ready to burst
Flower；
(2) pollen is dry: by the careful removal of petal of the flower of acquisition, holding chamber is interior to be spontaneously dried；When anther dehiscence loose powder, gently
Light vibration flower, pollen is equably collected on clean blank sheet of paper, continues to dry in the air dry to pollen, drying regime is that pollen is complete
It scatters, not at bulk；
(3) pollen storage: glass desicator internal washing is clean, and with 75% alcohol wipe inner wall, the silica gel that will change colour pours into
And it is covered with drier bottom；Dried pomegranate pollen is fitted into container, seals, is placed in the drier of sealing, in 4 DEG C
It saves.
2. pomegranate pollen storage practice according to claim 1, which is characterized in that live to the pomegranate pollen after storage
Power measurement, specific method: drop 1-2 drop of liquid culture is based on glass slide, takes the pomegranate pollen stored on a small quantity uniform with pin
It spreads in liquid medium, glass slide is placed in the culture dish that relative humidity is 85%-90%, 25 DEG C of conditions in incubator
Lower dark culture 12h, pollen germination.
3. pomegranate pollen storage practice according to claim 1, which is characterized in that in the step (1), full-bloom stage is four
The period that the four months bottoms in valley area are opened to early May, 70%~75% purple U.S. pomegranate flower.
4. pomegranate pollen storage practice according to claim 1, which is characterized in that in the step (1), reach maturity
Flower is the flower that flower ovary is obviously expanded, flower pattern is calabash shaped or tubular, and selected flower is complete flower.
5. pomegranate pollen storage practice according to claim 1, which is characterized in that in the step (2), natural drying is
It is dried for 24 hours under 25 DEG C of Indoor Natural illumination.
6. pomegranate pollen storage practice according to claim 1, which is characterized in that in the step (3), container 10mL
Centrifuge tube, preservation are placed in 4 DEG C of fridge freshness retaining layer storages.
7. pomegranate pollen storage practice according to claim 2, which is characterized in that the fluid nutrient medium is MS+
4.2g/L sucrose.
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| CN201811214967.4A CN109287623A (en) | 2018-10-18 | 2018-10-18 | A kind of purple U.S. pomegranate pollen storage practice |
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| CN201811214967.4A CN109287623A (en) | 2018-10-18 | 2018-10-18 | A kind of purple U.S. pomegranate pollen storage practice |
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| CN109964811A (en) * | 2019-04-29 | 2019-07-05 | 南京林业大学 | A kind of cross breeding method of the quick emasculation of pomegranate |
| CN113465298A (en) * | 2021-07-16 | 2021-10-01 | 四川农业大学 | South Asia loquat pollen drying and storing method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109964811A (en) * | 2019-04-29 | 2019-07-05 | 南京林业大学 | A kind of cross breeding method of the quick emasculation of pomegranate |
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| CN113465298A (en) * | 2021-07-16 | 2021-10-01 | 四川农业大学 | South Asia loquat pollen drying and storing method |
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Application publication date: 20190201 |